Chapter 24 Postembedding Labeling on Lowicryl K4M Tissue Sections: Detection and Modification of Cellular Components

1989 ◽  
pp. 513-551 ◽  
Author(s):  
J. Roth
Keyword(s):  
Tissue Sections ◽  
Lowicryl K4m ◽  
Cellular Components

Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells

1987 ◽  
Vol 45 ◽  
pp. 906-907
Author(s):  
W. E. Rigsby ◽  
D. M. Hinton ◽  
V. J. Hurst ◽  
P. C. McCaskey
Keyword(s):  
Polarized Light ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Whole Cells ◽  
Tissue Sections ◽  
Hepatic Cells ◽  
Slaughter House ◽  
Mammalian Tissues ◽  
Carbon Coated ◽  
Cellular Components

Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

scholarly journals Light and electron microscopic demonstration of sialic acid residues with the lectin from Limax flavus: a cytochemical affinity technique with the use of fetuin-gold complexes.

1984 ◽  
Vol 32 (11) ◽  
pp. 1167-1176 ◽  
Author(s):  
J Roth ◽  
J M Lucocq ◽  
P M Charest
Keyword(s):  
Electron Microscopy ◽  
Sialic Acid ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
Gold Complexes ◽  
Thin Sections ◽  
Tissue Sections ◽  
Lowicryl K4m ◽  
Hydrolysis Of ◽  
Electron Microscopic Demonstration

The development of a cytochemical affinity technique for the demonstration of sialic acid residues by light and electron microscopy is reported. The lectin from the slug Limax flavus, with its narrow specificity for N-acetyl- and N-glycolylneuraminic acid, was applied to tissue sections. Subsequently fetuin-gold complexes were used to visualize the tissue-bound lectin. Different cytochemical controls, including sugar inhibition tests, neuraminidase digestion, the use of fetuin-gold complexes alone, or acid hydrolysis of sections, proved the specificity of the technique. Postembedding staining was performed on frozen, paraffin, or semithin resin sections for light microscopy and on thin sections from low temperature Lowicryl K4M-embedded material for electron microscopy. The distribution of sialic acid residues in rat pancreas, liver, and colonic mucosa was investigated.

scholarly journals Seq-Scope: Submicrometer-resolution spatial transcriptomics for single cell and subcellular studies

2021 ◽  
Author(s):  
Chun-Seok Cho ◽  
Jingyue Xi ◽  
Sung-Rye Park ◽  
Jer-En Hsu ◽  
Myungjin Kim ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Molecule ◽  
Single Cell Analysis ◽  
Solid Phase ◽  
Cell Types ◽  
Optical Microscope ◽  
Tissue Sections ◽  
Spatial Coordinates ◽  
Wide Group ◽  
Cellular Components

AbstractSpatial barcoding technologies have the potential to reveal histological details of transcriptomic profiles; however, they are currently limited by their low resolution. Here we report Seq-Scope, a spatial barcoding technology with a resolution almost comparable to an optical microscope. Seq-Scope is based on a solid-phase amplification of randomly barcoded single-molecule oligonucleotides using an Illumina sequencing-by-synthesis platform. The resulting clusters annotated with spatial coordinates are processed to expose RNA-capture moiety. These RNA-capturing barcoded clusters define the pixels of Seq-Scope that are approximately 0.5-1 μm apart from each other. From tissue sections, Seq-Scope visualizes spatial transcriptome heterogeneity at multiple histological scales, including tissue zonation according to the portal-central (liver), crypt-surface (colon) and inflammation-fibrosis (injured liver) axes, cellular components including single cell types and subtypes, and subcellular architectures of nucleus, cytoplasm and mitochondria. Seq-scope is quick, straightforward and easy-to-implement, and makes spatial single cell analysis accessible to a wide group of biomedical researchers.

scholarly journals Monoclonal antibody specific for the T-tubule of skeletal muscle.

1986 ◽  
Vol 34 (3) ◽  
pp. 347-355 ◽  
Author(s):  
N N Malouf ◽  
S Taylor ◽  
G Y Gillespie ◽  
J M Bynum ◽  
P E Wilson ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibody ◽  
Sarcoplasmic Reticulum ◽  
Surface Membrane ◽  
Immunoperoxidase Staining ◽  
Rabbit Muscle ◽  
Tissue Sections ◽  
Electron Immunocytochemistry ◽  
Lowicryl K4m ◽  
T System

Monoclonal antibodies were raised against a triad-enriched (sarcoplasmic reticulum-T-tubule complex) microsomal membrane fraction of rabbit skeletal muscle. The avidin-biotin complex (ABC) immunoperoxidase staining method was used to screen hybrid colonies. Positive antibodies exhibited a granular doublet pattern at the A-I junction, consistent with the location of triads in rabbit muscle. One monoclonal antibody, M171, was further characterized by ultrastructural and immunoadsorption techniques. Postembedding electron immunocytochemistry was performed on tissue sections embedded in Lowicryl K4M. Goat anti-mouse immunoglobulin absorbed to 10 nm colloidal gold particles was used as an ultrastructural label. In these studies, M171 recognized an epitope at the triads and at periodic openings along the plasmalemma. Immunoadsorption on protein transfers of isolated sarcoplasmic reticulum, surface membrane (plasmalemma and T-tubule), and triad-enriched fractions showed that M171 reacts with a surface membrane component. Taken together, these studies suggest that M171 recognizes an epitope associated with the T-tubule at the triad and at the "mouth" of the T-system at the plasmalemma.

An enhanced cellular organelle visualization procedure when staining for peroxisomes

1995 ◽  
Vol 53 ◽  
pp. 886-887
Author(s):  
J. W. Horn ◽  
B.J. Dovey-Hartman
Keyword(s):  
Tissue Architecture ◽  
Ultrastructural Examination ◽  
Fixed Tissue ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Visual Identification ◽  
Transmission Electron ◽  
Light Staining ◽  
Cellular Components

Reliable visual identification of peroxisomes is important in developmental, clinical, and investigational research. The current technique employed in most laboratories uses a specific electron dense label for the demonstration of peroxisomes by transmission electron microscopy by applying 3,3'- Diaminobenzidine Tetrahydrochloride (DAB) directly to freshly fixed tissue samples to react with endogenous peroxisomal catalase. After routine processing, ultrastructural examination of tissue sections is conducted either with light staining or without post-staining of grids. While peroxisomes are easily identified using this method, remaining tissue architecture is difficult to visualize due to the opacity of the tissue. Additionally, if grids are post-stained with heavy metal solutions, they must be modified to allow for enough staining to visualize cellular components without compromising the quality of the peroxisome label. We will describe a technique whereby DAB-reacted tissues are stained with a postfixative solution including potassium ferricyanide that imparts density to cell membranes and cellular components thereby enhancing identification and interpretion of data.

scholarly journals Specific binding of fibronectin--antifibronectin immune complexes to procollagen: a new pitfall in immunostaining.

1984 ◽  
Vol 98 (3) ◽  
pp. 1042-1047 ◽  
Author(s):  
J A McDonald ◽  
D G Kelley
Keyword(s):  
Monoclonal Antibodies ◽  
Immune Complexes ◽  
Culture Supernatant ◽  
Specific Binding ◽  
Lung Fibroblasts ◽  
Pericellular Matrix ◽  
Tissue Sections ◽  
Rat Tissue ◽  
Cellular Components ◽  
Serum Components

We observed intense intracellular immunofluorescence of rat lung fibroblasts stained with hybridoma culture supernatant containing monoclonal antibodies to human plasma fibronectin, but no pericellular matrix staining. Immunoprecipitation and absorption experiments revealed that this intracellular staining by hybridoma-conditioned medium was due to binding of fibronectin-antifibronectin immune complexes via the fibronectin to intracellular procollagen. The anomalous staining patterns we encountered were not revealed by the usual controls for immunohistochemical specificity, and also occurred in rat tissue sections. This general phenomena--binding of serum antigens present in hybridoma medium to cellular components--could in principle result in artifactual staining with monoclonal antibodies to other serum components, so investigators using monoclonal antibodies should be aware of this new artifact. Our results also demonstrate that fibronectin binds specifically to native procollagen. Monoclonal antibodies may be useful for studying fibronectin-procollagen and other macromolecular interactions.

scholarly journals Morphology and Anatomy of Leaf, Stem and Petiole of Luvunga crassifolia Tanaka (Rutaceae)

2021 ◽  
Vol 17 (6) ◽  
pp. 818-828
Author(s):  
Sugathini Shunmugam ◽  
Nur Syamilah Rosli ◽  
Sugumaran Manickam ◽  
Nur Fatihah Mohd Yusoff ◽  
Noorjahan Banu Alitheen ◽  
...  
Keyword(s):  
Plant Species ◽  
Cross Sections ◽  
Diagnostic Features ◽  
Tissue Sections ◽  
Leaf Surfaces ◽  
Calcium Oxalates ◽  
Secretory Cavities ◽  
Histological Method ◽  
Cellular Components ◽  
Histological Procedure

Luvunga crassifolia is an underutilized plant in the Citrus family. Other than brief morphological descriptions, there are no published reports on other identification features of this plant. Thus, the current study was aimed to investigate macroscopic and microscopic diagnostic features of L. crassifolia leaves, stems, and petioles. Macroscopic characterization, optimization of histological procedure, and histochemical analyses of differential stains were carried out on the leaves, stems, and petioles of L. crassifolia. The histological method was optimized by modifying the following parameters: number of fixation days, dehydration duration with degraded series of ethanol or butanol, clearing duration, and infiltration duration. After infiltration, embedding and sectioning of the tissues were performed. Histochemical analyses were carried out using differential stains to identify the cellular components in leaf, stem and, petiole tissue sections. This study showed that L. crassifolia leaves are amphistomatic. Pellucid dots were observed on both adaxial and abaxial leaf surfaces. Secretory cavities, xylem, phloem, and pericyclic fibers were found in the cross-sections of leaf, stem, and petiole. Calcium oxalates were present in the leaf and stem sections, while trichomes were detected in stem and petiole sections. The information obtained from this study will be helpful for the identification and future taxonomic-related studies of this plant species.

Freeze-Fracture Replication in the Ultrastructure of Blue-Green Algae

1967 ◽  
Vol 25 ◽  
pp. 74-75
Author(s):  
L. V. Leak
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Green Algae ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Electron Microscopic ◽  
Photosynthetic Membranes ◽  
Blue Green Algae ◽  
Cell Biol ◽  
Cellular Components

Electron microscopic observations of freeze-fracture replicas of Anabaena cells obtained by the procedures described by Bullivant and Ames (J. Cell Biol., 1966) indicate that the frozen cells are fractured in many different planes. This fracturing or cleaving along various planes allows one to gain a three dimensional relation of the cellular components as a result of such a manipulation. When replicas that are obtained by the freeze-fracture method are observed in the electron microscope, cross fractures of the cell wall and membranes that comprise the photosynthetic lamellae are apparent as demonstrated in Figures 1 & 2.A large portion of the Anabaena cell is composed of undulating layers of cytoplasm that are bounded by unit membranes that comprise the photosynthetic membranes. The adjoining layers of cytoplasm are closely apposed to each other to form the photosynthetic lamellae. Occassionally the adjacent layers of cytoplasm are separated by an interspace that may vary in widths of up to several 100 mu to form intralamellar vesicles.

Mitochondrial Reconstructions Based on Serial Thick Sections and HVEM

1975 ◽  
Vol 33 ◽  
pp. 286-287
Author(s):  
Jerome J. Paulin
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Posterior Pole ◽  
Dimensional Structure ◽  
Stereo Imaging ◽  
Thin Sections ◽  
Anatomical Features ◽  
The Past ◽  
Cellular Components ◽  
Mitochondrial Reticulum

Within the past decade it has become apparent that HVEM offers the biologist a means to explore the three-dimensional structure of cells and/or organelles. Stereo-imaging of thick sections (e.g. 0.25-10 μm) not only reveals anatomical features of cellular components, but also reduces errors of interpretation associated with overlap of structures seen in thick sections. Concomitant with stereo-imaging techniques conventional serial Sectioning methods developed with thin sections have been adopted to serial thick sections (≥ 0.25 μm). Three-dimensional reconstructions of the chondriome of several species of trypanosomatid flagellates have been made from tracings of mitochondrial profiles on cellulose acetate sheets. The sheets are flooded with acetone, gluing them together, and the model sawed from the composite and redrawn.The extensive mitochondrial reticulum can be seen in consecutive thick sections of (0.25 μm thick) Crithidia fasciculata (Figs. 1-2). Profiles of the mitochondrion are distinguishable from the anterior apex of the cell (small arrow, Fig. 1) to the posterior pole (small arrow, Fig. 2).

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